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M-T7 modulates the immune response in the healing wound. (A) ELISA quantification of CCL2 in wounds treated with saline or M-T7 at days 1, 4 and 7 post-wounding, normalized to total protein. (B) Quantification of Arginase-1+ cells per 20× field of wounds treated with saline or M-T7 at days 2, 4 and 7 post-wounding. (C) Representative Arginase-1 IHC fields at day 7. (D) Quantification of TGF-beta+ cells per 20× field on days 2, 4 and 7 post-wounding. (E,F) Quantification of CD3+ cells per 20× field of wounds treated with saline or M-T7 at days 2, 4 and 7 post-wounding, specifically in the (E) wound bed or (F) epithelial tongue. (G) Quantification of CD4+ cells per 20× field of wounds treated with saline or M-T7 at days 2, 4 and 7 post-wounding normalized to the numbers on day 2. (H) Representative CD4 IHC fields in the epithelial tongue at day 7. Full 20× field is given in Figure S2. All bars are mean and standard error. Statistics are calculated by two-way ANOVA with Fisher's LSD post-hoc analysis. N = 3-4 in each group and time point.
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Complex dermal wounds represent major medical and financial burdens, especially in the context of comorbidities such as diabetes, infection and advanced age. New approaches to accelerate and improve, or "fine tune" the healing process, so as to improve the quality of cutaneous wound healing and management, are the focus of intense investigation. He...
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... also called monocyte chemoattractant protein-1 (MCP-1), is a CC-class chemokine previously shown to have a critical role in the regulation of wound healing [48]. We performed the ELISA analysis of CCL2 on wound tissues treated with saline or M-T7, collected on days 1, 4 and 7 post-wounding ( Figure 4A). Wounds treated with M-T7 had an elevated level of CCL2 on day 4 post-wounding, which approached significance (p = 0.0763), while levels of CCL2 were not different between saline and M-T7 treatment on days 1 and 7 post-wounding. ...
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... performed immunohistochemistry of wounds treated with saline or M-T7 on days 2, 4 and 7 post-wounding, staining for Arginase-1, a canonical marker of M2 macrophage polarization. The quantification of Arginase-1+ cells revealed a trend towards elevated M2 macrophages on days 2 and 4 post-wounding, which achieved significance (p < 0.05) by day 7 post-wounding ( Figure 4B,C). Accordingly, the number of TGF-beta+ cells per field trended towards significance on day 4 (p = 0.0836) and reached significance by day 7 post-wounding (p < 0.05) ( Figure 4D). ...
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... quantification of Arginase-1+ cells revealed a trend towards elevated M2 macrophages on days 2 and 4 post-wounding, which achieved significance (p < 0.05) by day 7 post-wounding ( Figure 4B,C). Accordingly, the number of TGF-beta+ cells per field trended towards significance on day 4 (p = 0.0836) and reached significance by day 7 post-wounding (p < 0.05) ( Figure 4D). We further investigated the effects of M-T7 treatment on T cell infiltration in the healing wound. ...
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... further investigated the effects of M-T7 treatment on T cell infiltration in the healing wound. We found that M-T7 treatment significantly inhibited the infiltration of CD3+ T cells, a general T cell marker, into the bed of the healing wound on days 4 and 7 post-wounding ( Figure 4E), without inhibiting the accumulation of CD3+ cells in the epithelial tongue of the wounds ( Figure 4F). Regulatory T cells, a CD4 T cell subtype, are crucial for the normal and accelerated healing of cutaneous wounds [50]. ...
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... further investigated the effects of M-T7 treatment on T cell infiltration in the healing wound. We found that M-T7 treatment significantly inhibited the infiltration of CD3+ T cells, a general T cell marker, into the bed of the healing wound on days 4 and 7 post-wounding ( Figure 4E), without inhibiting the accumulation of CD3+ cells in the epithelial tongue of the wounds ( Figure 4F). Regulatory T cells, a CD4 T cell subtype, are crucial for the normal and accelerated healing of cutaneous wounds [50]. ...
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... the dimerization of CCL2 requires glycosaminoglycan interactions [71]. We found increased levels of CCL2 in healing wounds when treated with recombinant M-T7 ( Figure 4A), consistent with the role of CCL2 in improved healing. We hypothesize that M-T7 treatment inhibited the oligomerization of CCL2, slowing receptor engagement and delaying its subsequent degradation. ...
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... further, we found M-T7-dependent effects in two cell populations known to be affected by CCL2 and other chemokine signaling: macrophages and T cells. Specifically, we found an increase in M2-polarized, proresolution macrophages ( Figure 4B). This finding agrees with prior work showing that CCL2 signaling results in M2 polarization [49]. ...
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... finding agrees with prior work showing that CCL2 signaling results in M2 polarization [49]. We also found increased CD4+ T cells in the epithelial tongues of healing wounds treated with M-T7 ( Figure 4F,G), in agreement with the ability for CCL2 to promote CD4 recruitment, and in the CD4-lineage cells driving accelerated wound healing [50,72]. CCL2 acts directly on T cells via the action of CCR2 and CCR4 [73], but can also induce the recruitment of CD4 cells into tissues in a promiscuous manner, using other receptors [72]. ...
Citations
... To combat ECTV infection, murine macrophages mediate T1IFN production in a cGAS-STING-dependent pathway [86] and are considered a good model for the study of ECTV immunopathogenesis [126]. MYXV also encodes for M− T7 and Serp-1, which do fine in regulating macrophages in different contexts, e.g., by decreasing macrophages infiltration in the cavity of injury in animals after spinal cord injury and attenuating inflammation [127,128], and also by increasing arginase-expressing macrophages and enhancing wound healing [129]. Recombinant VACV could be planned to prime macrophages to elicit such functions as antitumor immune responses [130]. ...
Poxviruses (PXVs) are mostly known for the variola virus, being the cause of smallpox; however, re-emerging PXVs have also shown a great capacity to develop outbreaks of pox-like infections in humans. The situation is alarming; PXV outbreaks have been involving both endemic and non-endemic areas in recent decades. Stopped smallpox vaccination is a reason offered mainly for this changing epidemiology that implies the protective role of immunity in the pathology of PXV infections. The immune system recognizes PXVs and elicits responses, but PXVs can antagonize these responses. Here, we briefly review the immunology of PXV infections, with emphasis on the role of pattern-recognition receptors, macrophages, and natural killer cells in the early response to PXV infections and PXVs’ strategies influencing these responses, as well as taking a glance at other immune cells, which discussion over them mainly occurs in association with PXV immunization rather than PXV infection. Throughout the review, numerous evasion mechanisms are highlighted, which might have implications for designing specific immunotherapies for PXV in the future.
... 6,7,114,115 Neutrophils, macrophages, B lymphocytes and T lymphocytes are active in the wound to establish an immune barrier against microbial invasion during this period. 116,117 However, the prolonged presence of neutrophils with excessive infiltration and inappropriate termination ways and the production of inflammatory mediators. 118 miRNA-126 and miRNA-23 overexpression, and miRNA-21 and miRNA-155 downregulation are involved in inflammatory mechanisms through phosphatidylinositide 3-kinases (PI3K)/protein kinase B (AKT)1/nuclear factor kappa-B (NF-kB) gene expression. ...
Skin wounds are a common condition causing economic burden and they represent an urgent clinical need, especially chronic wounds. Numerous studies have been conducted on the applications of stem cell therapy in wound healing, with adipose-derived mesenchymal stem cells (ADMSCs) playing a major role since they can be isolated easily, yielding a high number of cells, the less invasive harvesting required, the longer life span and no ethical issues. However, the lack of standardized doses and protocols, the heterogeneity of clinical trials, as well as the incompatibility of the immune system limit its application. Recent studies have demonstrated that specific stem cell functions depend on paracrine factors, including extracellular vesicles, in which microRNAs in exosomes (Exo-miRNAs) are essential in controlling their functions. This paper describes the application and mechanism whereby ADMSC-Exo-miRNA regulates wound healing. ADMSC-Exo-miRNA is involved in various stages in wounds, including modulating the immune response and inflammation, accelerating skin cell proliferation and epithelialization, promoting vascular repair, and regulating collagen remodeling thereby reducing scar formation. In summary, this acellular therapy based on ADMSC-Exo-miRNA has considerable clinical potential, and provides reference values for developing new treatment strategies for wound healing.
... As central immune organs in teleost fish, the spleen and head kidney are responsible for regulating immune response (8), containing large numbers of lymphocytes and macrophages (9,10). As we all know, lymphocytes and macrophages are an essential part of the immune system (11). The fish's skin, an important mucosal defense organ, has developed a mature immune barrier to protect the whole body from pathogens invasion (12,13). ...
Aflatoxin B1 (AFB1) is kind of a common mycotoxin in food and feedstuff. Aquafeeds are susceptible to contamination of AFB1. In teleost fish, the spleen and head kidney are key immune organ. Moreover, the fish skin is a critical mucosal barrier system. However, there was little study on the effects of dietary AFB1 on the immune response of these immune organs in fish. This study aimed to explore the impacts of oral AFB1 on the immune competence and its mechanisms in the skin, spleen, and head kidney of grass carp. Our work indicated that dietary AFB1 reduced antibacterial compounds and immunoglobulins contents, and decreased the transcription levels of antimicrobial peptides in grass carp immune organs. In addition, dietary AFB1 increased the transcription levels of pro-inflammatory cytokines and reduced the transcription levels of anti-inflammatory cytokines in the grass carp immune organs, which might be regulated by NF-κB and TOR signaling, respectively. Meanwhile, we evaluated the content of AFB1 in the grass carp diet should not exceed 29.48 μg/kg diet according to the levels of acid phosphatase and lysozyme. In summary, dietary AFB1 impaired immune response in grass carp skin, spleen, and head kidney.
... The splinted full thickness model was used study the effects of recombinant myxoma virus-derived immune modulator M-T7 [49], of Babassu oil [50], of three-dimensional printed scaffolds and electrospun mats [51], of adult gingival multipotent mensenchymal stem cells [52], of induced pluripotent stem cells clay [53], of poly(lactic-co-glycolic acid) and vascular endothelial growth factor [54], and of renal dysfunction [44] on cutaneous wound healing. ...
A large number of models are now available for the investigation of skin wound healing. These can be used to study the processes that take place in a phase-specific manner under both physiological and pathological conditions. Most models focus on wound closure, which is a crucial parameter for wound healing. However, vascular supply plays an equally important role and corresponding models for selective or parallel investigation of microcirculation regeneration and angiogenesis are also described. In this review article, we therefore focus on the different levels of investigation of skin wound healing (in vivo to in virtuo) and the investigation of angiogenesis and its parameters.
... This finding is entirely consistent with prior research both in pre-clinical or clinical studies wherein Serp-1 demonstrated no significant toxicity and no neutralizing antibodies were detected in the Phase Further studies should also aim to investigate the molecular mechanism of action of Serp-1 in its anti-inflammatory role. Finally, a recent publication from our group demonstrated that administration of recombinant M-T7 protein (another Myxoma virus protein) could also accelerate dermal wound healing (24). This provides an additional example of a Myxoma virus derived protein in facilitating tissue repair. ...
Purpose: Chemical corneal injuries carry a high morbidity and commonly lead to visual impairment. Here, we investigate the role of Serp-1, a serine protease inhibitor, in corneal wound healing.
Methods: An alkaline-induced corneal injury was induced in 14 mice. Following injury, five mice received daily topical saline application while nine mice received Serp-1 100 μL topically combined with a daily subcutaneous injection of 100 ng/gram body weight of Serp-1. Corneal damage was monitored daily through fluorescein staining and imaging. Cross sectional corneal H&E staining were obtained. CD31 was used as marker for neovascularization.
Results: Serp-1 facilitates corneal wound healing by reducing fibrosis and neovascularization while mitigating inflammatory cell infiltration with no noticeable harm related to its application.
Conclusions: Serp-1 effectively mitigates inflammation, decreases fibrosis, and reduce neovascularization in a murine model of corneal injury without affecting other organs.
Translational Relavence: Our study provides preclinical data for topical application of Serp-1 to treat corneal wounds.
In this study, a new wound dressing was developed to speed up the healing process of diabetic wounds. First of all, taxifolin liposome (TL) was manufactured in this study. Then, taxifolin (TAX) and TL were mixed with polyvinyl alcohol (PVA) and chitosan (CS) by electrostatic spinning to prepare nanocomposite membranes. Finally, the mechanism of nanocomposite membranes to accelerate diabetic wound healing was investigated. The diameter of TL-loaded polyvinyl alcohol/chitosan nanocomposite membranes (PVA/CS/TL) was 429.43 ± 78.07 nm. The results of in vitro experiments demonstrated that the PVA/CS/TL had better water absorption, water vapor transmission rate (WVTR), porosity, hydrophilicity, mechanical properties, slow-release, antioxidant capacity, and antibacterial properties. The results of in vivo experiments demonstrated that the wound healing rate of mice treated with PVA/CS/TL for eighteen days was 98.39 ± 0.34 %. Histopathological staining, immunohistochemical staining, and western blot experiments also demonstrated that PVA/CS/TL could promote wound healing in diabetic mice by inhibiting the activation of inhibitor kappa B alpha (IκBα)/nuclear factor-kappa B (NF-κB) signaling pathway and related pro-inflammatory factors to increase the expression of CD31 and VEGF in skin tissues. These results suggested that PVA/CS/TL could be a potential candidate for wound dressing to promote chronic skin wound healing.
Growth factor therapy has demonstrated great promise for chronic wound repair, but controlling growth factor activity and cell phenotype over desired time frames remains a critical challenge. In this study, we developed a gene-activated hyaluronic acid-collagen matrix (GAHCM) comprising DNA/polyethylenimine (PEI) polyplexes retained on hyaluronic acid (HA)-collagen hydrogels using collagen mimetic peptides (CMPs). We hypothesized that manipulating both the number of CMP-collagen tethers and the ECM composition would provide a powerful strategy to control growth factor gene transfer kinetics while regulating cell behavior, resulting in enhanced growth factor activity for wound repair. We observed that polyplexes with 50% CMP-modified PEI (50 CP) showed enhanced retention of polyplexes in HCM hydrogels by 2.7-fold as compared to non-CMP modified polyplexes. Moreover, the incorporation of HA in the hydrogel promoted a significant increase in gene transfection efficiency based upon analysis of Gaussia luciferase (GLuc) reporter gene expression, and gene expression could be attenuated by blocking HA-CD44 signaling. Furthermore, when fibroblasts were exposed to vascular endothelial growth factor-A (VEGF-A)-GAHCM, the 50 CP matrix facilitated sustained VEGF-A production for up to 7 days, with maximal expression at day 5. Application of these VEGF-A-50 CP samples stimulated prolonged pro-healing responses, including the TGF-β1-induced myofibroblast-like phenotypes and enhanced closure of murine splinted wounds. Overall, these findings demonstrate the use of ECM-based materials to stimulate efficient gene transfer and regulate cellular phenotype, resulting in improved control of growth factor activity for wound repair. GAHCM have significant potential to overcome key challenges in growth factor therapy for regenerative medicine.
Statement of Significance
Despite great promise for growth factor therapies in wound treatment, controlling growth factor activity and providing a microenvironment for cells that maximizes growth factor signaling have continued to limit the success of existing formulations. Our GAHCM strategy, combining CMP gene delivery and hyaluronic acid-collagen matrix, enabled enhanced wound healing efficacy via the combination of controlled and localized growth factor expression and matrix-mediated regulation of cell behavior. Incorporation of CMPs and HA in the same matrix synergistically enhanced VEGF activity as compared with simpler matrices. Accordingly, GAHCM will advance our ability to leverage growth factor signaling for wound healing, resulting in new long-term treatments for recalcitrant wounds.
Bacterial infection is a major obstacle to the wound healing process. The hydrogel dressings with a simpler structure and good antibacterial and wound healing performance are appealing for clinical application. Herein, a robust hydrogel was synthesized from acrylamide (AM), acrylic acid (AA) and N,N′-methylene diacrylamide (MBA) via a redox initiating polymerization. The polymerization conditions were optimized to obtain the hydrogel with minimum unreacted monomers, which were 0.25% and 0.12% for AM and AA, respectively. The hydrogel had good mechanical strength, and could effectively resist damage by external forces and maintain a good macroscopic shape. It showed large water uptake capacity, and could post load a wide range of molecules via hydrogen bonding and electrostatic interaction. Loading of antibiotic doxycycline (DOX) enabled the hydrogel with good antibacterial activity against both Gram-positive bacteria and Gram-negative bacteria in vitro and in vivo. In a rat model of methicillin-resistant Staphylococcus aureus (MRSA)-infected full-thickness skin defect wound, the DOX-loaded hydrogel showed good therapeutic effect. It could significantly promote the wound closure, increased the collagen coverage area, down-regulate the expressions of pro-inflammatory TNF-α and IL-1β factors, and up-regulate the expressions of anti-inflammatory IL-4 factor and CD31 neovascularization factor.
